cmet  (R&D Systems)

Journal: mAbs

Article Title: Modular generation of multispecific antibodies using protein trans-splicing

doi: 10.1080/19420862.2025.2573179

Figure Lengend Snippet: Monovalent bispecific IgGs produced by protein splicing recapitulate the functional properties of DuetMab benchmarks in cellular assays. (A) Cartoon representation of the six monovalent bispecific antibodies generated by protein splicing. Each cartoon is positioned directly above the assays in which the molecule was evaluated in subsequent panels. (B) Cellular binding of spliced bispecific antibodies and their DuetMab benchmarks to EGFR + /HER2 + /cMET + NCI-H358 cells and (C) CD3 + HPB-ALL cells. (D) direct, (E) ADC cellular cytotoxicity, and (F) T-cell engager cytotoxicity assays comparing spliced molecules to their DuetMab benchmarks. Nip228 is a non-binding isotype control antibody. Cell binding curves are representative examples from n = 2 experiments. Values for direct, ADC, TCE cytotoxicity represent the mean ± the standard error about the mean of two biological replicates.

Article Snippet: The EGFR+, HER2+, and cMET+ cell line NCI H358 were obtained from the American Type Culture Collection (cat# CRL-5807), and CD3+ cell line HPB-ALL was obtained from DSMZ (cat # ACC 483).

Techniques: Produced, Functional Assay, Generated, Binding Assay, Control

Journal: mAbs

Article Title: Modular generation of multispecific antibodies using protein trans-splicing

doi: 10.1080/19420862.2025.2573179

Figure Lengend Snippet: Production and characterization of monovalent trispecific antibodies by protein splicing. (A) Protein trans-splicing schematic in which an αEGFR-αCD3 Int C -scaffolds and an αHER2 Fab-Int N are reacted to produce a monovalent trispecific antibodies with two distinct geometries. (B) SDS-PAGE analysis of the intein reactants and trispecific antibody products (i, light chain; ii, Fab heavy chain; iii, Int C heavy chain; iv, Fab-Int N ; v, spliced heavy chain). (C) Analytical SEC of the spliced trispecific antibodies and a monovalent bispecific DuetMab. (D) EGFR, HER2, and CD3 antigen binding of the spliced trispecifics compared to monovalent bispecific DuetMab controls measured by biolayer interferometry. (E) Cellular binding of spliced trispecific antibodies to EGFR + /HER2 + NCI-H358 cells and CD3 + HPB-ALL cells. (F) T-cell engager cytotoxicity assays comparing activity of the two geometric orientations of the spliced trispecific antibodies.. Cell binding curves are representative examples from n = 2 experiments. SDS-PAGE, SEC, and BLI measurements are performed post post Ni-NTA cleanup and are representative examples from n = 2 replicates. Values for direct and TCE cytotoxicity represent the mean ± the standard error about the mean of two biological replicates.

Article Snippet: The EGFR+, HER2+, and cMET+ cell line NCI H358 were obtained from the American Type Culture Collection (cat# CRL-5807), and CD3+ cell line HPB-ALL was obtained from DSMZ (cat # ACC 483).

Techniques: SDS Page, Binding Assay, Activity Assay

Journal: mAbs

Article Title: Modular generation of multispecific antibodies using protein trans-splicing

doi: 10.1080/19420862.2025.2573179

Figure Lengend Snippet: Function of bivalent multispecific antibodies produced via dual-splicing. (A) Cartoon representation of the generated multispecific antibodies. Each cartoon is positioned directly above the assays in which the molecule was evaluated in subsequent panels. (B) Cellular binding of spliced multispecific antibodies to EGFR + /HER2 + NCI-H358 cells. (C) Direct and (D) ADC cytotoxicity assays of spliced molecules. (E) Cellular binding to CD3 + HPB-ALL cells. (F) T-cell engager cytotoxicity assay of spliced molecules. Nip228 is a non-binding isotype control antibody. Values for direct cytotoxicity and TCE cytotoxicity represent the mean ± the standard error about the mean of two biological replicates. Values for ADC cytotoxicity represent the mean ± the standard error about the mean of four biological replicates.

Article Snippet: The EGFR+, HER2+, and cMET+ cell line NCI H358 were obtained from the American Type Culture Collection (cat# CRL-5807), and CD3+ cell line HPB-ALL was obtained from DSMZ (cat # ACC 483).

Techniques: Produced, Generated, Binding Assay, Cytotoxicity Assay, Control

Journal: Cell Reports Medicine

Article Title: Development of a cyst-targeted therapy for polycystic kidney disease using an antagonistic dimeric IgA monoclonal antibody against cMET

doi: 10.1016/j.xcrm.2025.102335

Figure Lengend Snippet: Recombinant cMET-dIgA displays potent antagonist activity and retains the ability to transcytose via pIgR (A) cMET-dIgA specifically bound mouse (top row), human (2 nd row), and rat (3rd row) cMET in transiently transfected CHO cells. An empty DNA vector (bottom row) was used for control. Human dIgA (used as a primary detection antibody, green); cMET (commercial IgG specific for all 3 species, red); DAPI (blue). Scale bar, 20μM. (B) cMET-dIgA blocked endogenous cMET receptor activation in mouse IMCD-3 cells after HGF stimulation (left panels) and did not cause receptor dimerization in the absence of HGF (right panels). Capmatinib (cMET SM inhibitor) was used as a negative control. Cells were lysed in SDS and analyzed via immunoblot. Quantification of immunoblot signal intensity from +HGF/+dIgA samples was performed using Image Studio Lite, normalized to actin. See also . (C) Transcytosis of cMET-dIgA visualized via immunoblot of human IgA in apical or basolateral (BL) media samples from MDCK cells (with/without pIgR stably transfected). All samples were run under nonreducing conditions. A vertical line between lanes indicates that the two sections of the blot were adjusted separately on Photoshop to improve visibility. (D) Transcytosis into apical compartments also occurred under low-calcium conditions in MDCK cells expressing pIgR. GP-135 (apical membrane marker, red); human dIgA (used as a primary detection antibody, green); DAPI (blue). Scale bar, 10 μM.

Article Snippet: cMET c-terminal , ProSci , Cat#79-590; RRID: AB_2332792.

Techniques: Recombinant, Activity Assay, Transfection, Plasmid Preparation, Control, Activation Assay, Negative Control, Western Blot, Stable Transfection, Expressing, Membrane, Marker

Journal: Cell Reports Medicine

Article Title: Development of a cyst-targeted therapy for polycystic kidney disease using an antagonistic dimeric IgA monoclonal antibody against cMET

doi: 10.1016/j.xcrm.2025.102335

Figure Lengend Snippet: cMET-dIgA traffics to PKD Han rat cysts and downregulates cMET signaling in cyst-lining cells (A and B) Untreated PKD Han:SPRD adult male rats upregulate pIgR expression compared to WT as seen in (A) immunoblot of whole kidney lysates, n = 3, as well as (B) paraffin-embedded kidney sections stained by immunofluorescence for pIgR (green), AQP1 (red), and DAPI (blue). Rats were injected with a single dose of 1 mg cMET-dIgA and sacrificed 24 h later, n = 5. (C) Immunoprecipitation of CF using nitrilotriacetic acid (NTA)-nickel (Ni) ion resin followed by immunoblot specific for human IgA1 revealed secretory IgA (sIgA) recovery. Un-injected purified dimeric IgA (dIgA) was loaded for size comparison. A vertical line between lanes indicates that the two sections of the blot were adjusted separately on Photoshop to improve visibility. (D) Immunofluorescent staining of C-terminal cMET (red), AQP1 (red), and DAPI (blue), as well as immunohistochemistry staining of phospho-cMET, decreased after dIgA administration. (E) Immunofluorescent staining of Ki67 (cyan) and DAPI (red) showed the most significant decrease in cyst-lining cells (white arrowheads), with quantification of the fraction of Ki67+ nuclei in a cyst. Each point on the graph represents that percentage for a single cyst in a given field. All image scale bars, 100 μm. Statistical analyses were performed using Mann-Whitney unpaired one-tailed t test and represented as mean ± SEM. A p value of less than 0.05 was considered significant.

Article Snippet: cMET c-terminal , ProSci , Cat#79-590; RRID: AB_2332792.

Techniques: Expressing, Western Blot, Staining, Immunofluorescence, Injection, Immunoprecipitation, Purification, Comparison, Immunohistochemistry, MANN-WHITNEY, One-tailed Test

Journal: Cell Reports Medicine

Article Title: Development of a cyst-targeted therapy for polycystic kidney disease using an antagonistic dimeric IgA monoclonal antibody against cMET

doi: 10.1016/j.xcrm.2025.102335

Figure Lengend Snippet: cMET-dIgA accumulates and stabilizes in Bpk mouse cyst lumen via pIgR-mediated transport (A) Untreated Bpk mice, P14, upregulate pIgR expression in whole kidney lysates compared to age-matched WT mice. (B) Serum (S), CF-depleted kidneys (DKs), and cyst fluid (CF) from Juvenile Bpk mice were analyzed 1, 3, 5, and 8 days post-dIgA injection. Sample concentrations were equalized via spectrophotometry. See also . (C) Recovery of dIgA in serum and CF was quantified 1 and 3 days post-injection via sandwich ELISA. Triplicate values from each +dIgA timepoint group (n = 2) was plotted from CF or serum and represented as mean ± SEM. (D) Side-by-side comparison of these samples on a nonreducing SDS-PAGE gel. (E) Immunoprecipitation of CF using NTA-Ni resin and antibodies specific for either human IgA1 (nonreduced) or secretory component (SC) (reduced). Un-injected cMET-dIgA was loaded on all immunoblots for size comparison of dimeric (d) and secretory (s) IgA. A vertical line between lanes indicates that the two sections of the blot were adjusted separately on Photoshop to improve visibility. (F) Immunofluorescent staining of human IgA (green) and DAPI (blue) shows targeting of cMET-dIgA to renal cyst lumen. Scale bar, 100 μm.

Article Snippet: cMET c-terminal , ProSci , Cat#79-590; RRID: AB_2332792.

Techniques: Expressing, Injection, Spectrophotometry, Sandwich ELISA, Comparison, SDS Page, Immunoprecipitation, Western Blot, Staining

Journal: Cell Reports Medicine

Article Title: Development of a cyst-targeted therapy for polycystic kidney disease using an antagonistic dimeric IgA monoclonal antibody against cMET

doi: 10.1016/j.xcrm.2025.102335

Figure Lengend Snippet: cMET-dIgA treatment causes potent target pathway inhibition in rapid-onset Bpk mouse model (A) Short-term treatment timeline in Bpk model that begins cystogenesis in utero (ED). WT and Bpk mice were injected with 20 mg/kg/day cMET-dIgA or vehicle for 1 week starting at post-natal day (P)7. Analysis of kidney sections from these mice included (B) immunofluorescent staining for total cMET (red), proximal tubules stained with LTL ( Lotus tetragonolobus lectin) marker (green), and DAPI (blue); (C) quantification of total cMET from (B) in cyst-lining cells; (D) immunohistochemistry stain of phospho-cMET; (E) immunofluorescent staining for phospho-ERK1/2 (red), human IgA (green), and DAPI (blue); (F) quantification of phospho-ERK from (E) in cyst-lining cells; and (G) immunohistochemistry stain of phospho-AMPK. Nuclei were counterstained with hematoxylin. All image scale bars, 50 μm. Statistical analyses were performed using Mann-Whitney unpaired one-tailed t test and represented as mean ± SEM. A p value of less than 0.05 was considered significant.

Article Snippet: cMET c-terminal , ProSci , Cat#79-590; RRID: AB_2332792.

Techniques: Inhibition, In Utero, Injection, Staining, Marker, Immunohistochemistry, MANN-WHITNEY, One-tailed Test

Journal: Cell Reports Medicine

Article Title: Development of a cyst-targeted therapy for polycystic kidney disease using an antagonistic dimeric IgA monoclonal antibody against cMET

doi: 10.1016/j.xcrm.2025.102335

Figure Lengend Snippet: cMET-dIgA treatment slows disease progression in rapid-onset Bpk mouse model H&E stain of Bpk kidney sections after 1 week of daily i.p cMET-dIgA or vehicle injections. (A–F) (A) Representative images, scale bar = 100 μm, and (B) stitched composite images, scale bar, 1 mm. Quantification of (C) cystic index by Photoshop grid scoring, (D) cortical cyst area as a fraction of the total tissue area using ImageJ, (E) two-kidney-to-body weights (2K/BW) of mice from each group after treatment, and (F) creatinine concentrations in BPK serum samples using QuantiChrom creatinine assay kit. (G–J) (G) Immunofluorescent staining for the proliferation marker, Ki67 (cyan) and DAPI (red); scale bar, 50 μm; (H) TUNEL stain for apoptotic cells (cyan) and DAPI (red); scale bar, 100 μm, with quantification of the fraction of Ki67+ or TUNEL+ nuclei in a Bpk cyst or in WT tubular epithelial cells. Each point on the graph represents that percentage for a single cyst or tubule in a given field. dIgA treatment had no significant effect on (I) body weight or (J) lung weight as a fraction of total body weight compared to vehicle treatment in WT or Bpk mice. Statistical analyses were performed using Mann-Whitney unpaired one-tailed t test and represented as mean ± SEM. A p value of less than 0.05 was considered significant.

Article Snippet: cMET c-terminal , ProSci , Cat#79-590; RRID: AB_2332792.

Techniques: Biomarker Discovery, Staining, Marker, TUNEL Assay, MANN-WHITNEY, One-tailed Test

Journal: Cell Reports Medicine

Article Title: Development of a cyst-targeted therapy for polycystic kidney disease using an antagonistic dimeric IgA monoclonal antibody against cMET

doi: 10.1016/j.xcrm.2025.102335

Figure Lengend Snippet: cMET-dIgA treatment prevents cyst expansion in an orthologous Pkd1 mouse model (A–G) (A) Treatment timeline in tamoxifen-inducible Pkd1 fl/fl mouse model. PKD1 gene excision was induced on p10 followed by single i.p. injections of either 10 mg/kg/2 day cMET-dIgA or vehicle starting at 3 weeks post-induction (p31) for 2 weeks and sacrificed 2 days after the final injection (p45). Analysis of paraffin-embedded kidney sections from these mice ( n = 4) included immunofluorescent staining for (B) cMET or TUNEL (cyan) and DAPI (red), scale bar, 100um; quantification using ImageJ of (C) total cMET fluorescent intensity in cyst cells, median denoted in each group; and (D) TUNEL + cyst cells as a fraction of total nuclei in the cyst; (E) H&E stain-representative images; scale bar, 100 μm; stitched composite images, scale bar, 1 mm. (F) Two kidney weight as a fraction of total body weight (p45). (G) Quantification of individual cyst areas using ImageJ. Each point represents one cyst; median denoted in each group. (H–J) (H) Sirius Red Fast Green collagen (red) staining, scale bar, 100 μm; quantification (data not shown) was performed using Adobe Photoshop/grid scoring. Analysis of (I) serum creatinine and (J) blood urea nitrogen (BUN) in serum of WT and PKD (−/+dIgA) mice. (K) Concentrations of cMET-dIgA recovered on p45 in CF, serum, kidney lysate, and lung lysate via sandwich ELISA specific for human IgA1. See also and . Statistical analyses were performed using Mann-Whitney unpaired one-tailed t test and represented as mean ± SD. A p value of less than 0.05 was considered significant.

Article Snippet: cMET c-terminal , ProSci , Cat#79-590; RRID: AB_2332792.

Techniques: Injection, Staining, TUNEL Assay, Sandwich ELISA, MANN-WHITNEY, One-tailed Test